Abstract: The genome size varies greatly in higher plants. Repetitive sequences account for most of the large plant ge-nomes while low-copy or single copy genic sequences, referred to as gene space, take up only a small portion of the ge-nomes. Considering the large amount of repetitive sequences, it is a great challenge to obtain genic sequences using high-throughout methods in non-model plants bearing large genomes. Currently, several approaches have been developed for gene enrichment on a genome-wide scale, such as cDNA library, methylation filtration library, high Cot library and transposon tagging. Here, we reviewed the technical principles, advantages and disadvantages of these methods, as well as the recent development of methylation filtration technology. An in-depth discussion was performed for selection of one method or combination of methods according to the research objectives and plant materials, especially for plants with large genomes.